Article ; Online: A gas phase fractionation acquisition scheme integrating ion mobility for rapid diaPASEF library generation.
2023 Volume 23, Issue 7-8, Page(s) e2200038
Abstract: Data independent acquisition (DIA/SWATH) MS is a primary strategy in quantitative proteomics. diaPASEF is a recent adaptation using trapped ion mobility spectrometry (TIMS) to improve selectivity/sensitivity. Complex DIA spectra are typically analyzed ... ...
Abstract | Data independent acquisition (DIA/SWATH) MS is a primary strategy in quantitative proteomics. diaPASEF is a recent adaptation using trapped ion mobility spectrometry (TIMS) to improve selectivity/sensitivity. Complex DIA spectra are typically analyzed with reference to spectral libraries. The best-established method for generating libraries uses offline fractionation to increase depth of coverage. More recently strategies for spectral library generation based on gas phase fractionation (GPF), where a representative sample is injected serially using narrow DIA windows that cover different mass ranges of the complete precursor space, have been introduced that performed comparably to deep offline fractionation-based libraries. We investigated whether an analogous GPF-based approach that accounts for the ion mobility (IM) dimension is useful for the analysis of diaPASEF data. We developed a rapid library generation approach using an IM-GPF acquisition scheme in the m/z versus 1/K0 space requiring seven injections of a representative sample and compared this with libraries generated by direct deconvolution-based analysis of diaPASEF data or by deep offline fractionation. We found that library generation by IM-GPF outperformed direct library generation from diaPASEF and had performance approaching that of the deep library. This establishes the IM-GPF scheme as a pragmatic approach to rapid library generation for analysis of diaPASEF data. |
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MeSH term(s) | Proteomics/methods ; Peptide Library ; Chemical Fractionation/methods ; Proteome/analysis | ||||||||||
Chemical Substances | Peptide Library ; Proteome | ||||||||||
Language | English | ||||||||||
Publishing date | 2023-03-10 | ||||||||||
Publishing country | Germany | ||||||||||
Document type | Journal Article ; Research Support, Non-U.S. Gov't | ||||||||||
ZDB-ID | 2032093-0 | ||||||||||
ISSN | 1615-9861 ; 1615-9853 | ||||||||||
ISSN (online) | 1615-9861 | ||||||||||
ISSN | 1615-9853 | ||||||||||
DOI | 10.1002/pmic.202200038 | ||||||||||
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Database | MEDical Literature Analysis and Retrieval System OnLINE |
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